This is a strand-specific RNA library construction kit developed for Illumina's high-throughput sequencing platform. It is designed for RNA library construction of 100 pg ~ 100 ng RNA samples, and contains all the components required for RNA fragmentation, reverse transcription (RT), and PCR library amplification. The product is designed for RNA fragmentation by heating, and then reverse transcription with a 3′-N6 Primer with a splice sequence, which relies on the template-switching activity of the reverse transcriptase enzyme to add a splice sequence at the 5′ end of the RNA to produce a cDNA with splice sequences at both ends. AccuNext Sequencing libraries for the Illumina sequencing platform can be obtained by PCR amplification of CDI-joining primers (Illumina, for RNA libraries) (Code No. AG12505, AG12506, AG12507).
This product is easy to operate and can obtain RNA sequencing libraries without the traditional tedious steps of end repair and splice connection; the optimized reaction system improves the library conversion efficiency and is compatible with different template amounts, so that the data obtained from sequencing is highly uniform and has complete coverage; at the same time, the directionality of the template conversion reaction retains the strand directionality of the RNA, so that RNA strand-specific sequencing data can be obtained.
Since rRNA accounts for nearly 90% of the total RNA, it is recommended to remove the rRNA and then construct the RNA library to reduce the proportion of rRNA in the sequencing and obtain more effective data in order to obtain more effective data during the sequencing and to reduce the sequencing cost.
The reaction system of this product has been carefully optimized, please use the reagents provided in this product for RNA fragmentation, reverse transcription and PCR library amplification experiments, and it is recommended that you do not change the dosage and concentration of any of the reaction components, or replace the components in this product with other equivalent products to avoid obtaining bad results. If substitution is required, please verify first.
2. Wide range of templates: RNA library construction for 100 pg ~ 100 ng RNA; compatible with eukaryotic and prokaryotic RNA samples; degraded RNA can also be successfully constructed, and compatible with RNA samples with RIN values from 2 to 10.
3. Simple operation: RNA sequencing libraries can be obtained without the traditional tedious steps of end repair and junction connection.
4. Accurately recognizes RNA strand specificity: Read 1 reads the antisense strand sequence of the original RNA, and Read 2 reads the positive-sense strand sequence of the original RNA;
5. Stable product performance: Repeatable experimental operations.
Package 2-1 is composed as follows (stored at -80°C):
| individual parts making up a compound | AG12503 | AG12504 |
| Control Total RNA* (1 μg / μl) | 20 μl | 20 μl |
| 5′-Adapter Primer Mix | 54 μl | 216 μl |
*: Control Total RNA is Mouse Liver Total RNA.
Package 2-2 is composed as follows (stored at -20°C):
| individual parts making up a compound | AG12503 | AG12504 |
| RNase Inhibitor (40 U/μl) | 6 μl | 24 μl |
| 3′-N6 Primer | 12 μl | 48 μl |
| 5X First-Strand Buffer | 48 μl | 192 μl |
| AccuNext Reverse Transcriptase (100 U/μl) | 24 μl | 96 μl |
| 2X AccuNext PCR Mix II | 300 μl | 1.2 ml |
| Nuclease free water | 1 ml | 1 ml X 4 pcs |
Attention:AccuNext CDI Junction Primers (Illumina, for RNA libraries) (Code No. AG12505, AG12506, AG12507) are required for the experiment, but are not configured in this product and need to be purchased separately.AccuNext Information on CDI junction primers (Illumina, for RNA libraries) is provided in Appendices A and B.
Transportation Temperature: Package 2-1 Dry Ice Transportation Package 2-2 -20℃ Ice Pack Transportation or Dry Ice Transportation
